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目的探讨脊髓薄片器官型培养的方法及其腹角α运动神经元、背角中间神经元的鉴别。方法利用出生8d乳鼠的腰段脊髓组织切片建立脊髓器官型培养模型,并用神经元的特异性SMI蛳32和Calretinin单克隆抗体进行免疫组化染色,对脊髓腹角α运动神经元和背角中间神经元加以鉴定,测定培养液中乳酸脱氢酶(LDH)的含量。结果脊髓片在体外生长良好,形态完整,α运动神经元和背角中间神经元的数目及各时点培养液中LDH含量恒定,脊髓片可存活2个月以上。结论脊髓的器官培养技术为研究脊髓生理、病理改变及神经保护提供了有效的方法。
Objective To investigate the method of organotypic culture of spinal cord slices and to differentiate the α motor neurons and the dorsal horn interneurons in the ventral horn. Methods The lumbar spinal cord tissue sections of 8-day-old neonatal rats were used to establish the model of spinal cord organ culture. Immunohistochemical staining was performed with neuron-specific SMI-32 and Calretinin monoclonal antibodies. The effects of α-motor neuron and dorsal angle The interneurons were identified and the content of lactate dehydrogenase (LDH) in the culture medium was determined. Results The spinal cord slices grew well in vitro and the morphology was intact. The number of α-motor neurons and the dorsal horn interneurons and the content of LDH in each culture medium were constant, and the spinal cord slices could survive for more than 2 months. Conclusion The technique of organ culture in the spinal cord provides an effective method for the study of spinal cord physiology, pathological changes and neuroprotection.